Investigating the regulation and substrate processing of ADAMTS13 and ADAMTS7 metalloproteases.

研究 ADAMTS13 和 ADAMTS7 金属蛋白酶的调节和底物加工。

• 批准号: 10450886
• 负责人: Joshua Mutambuki Muia
• 金额: $ 33.35万
• 依托单位: OSU CENTER FOR HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10450886
• 关键词: ADAMTS; Address; Adopted; Allosteric Regulation; Atherosclerosis; Biochemical; Bioinformatics; Biological Process; Blood Coagulation Disorders; Blood coagulation; Cardiovascular Diseases; Cardiovascular system; Catalysis; Conflict (Psychology); Coronary Arteriosclerosis; Defect; Development; Diagnosis; Disease; Disease Outcome; Disintegrins; Elements; Enzymatic Biochemistry; Enzymes; Family; Foundations; Goals; Inflammation; Inflammatory; Inherited; Investigation; Kinetics; Literature; Metalloproteases; Methods; Modeling; Molecular Conformation; Outcome; Peptide Hydrolases; Prevention; Principal Investigator; Property; Proteomics; Reagent; Regulation; Reporting; Research; Thrombosis; Thrombospondins; Work; biochemical tools; human disease; improved; member; novel; protein protein interaction; recruit; small molecule inhibitor; student mentoring; therapeutic target; tool; translational applications; von Willebrand Factor

Project Summary This MIRA proposal aims to uncover the regulatory mechanisms and substrates for metalloproteases in the ADAMTS (A Disintegrin And Metalloprotease with ThromboSpondin motifs) family. These proteases are thought to be involved in many biological processes and some members are known to contribute to important human diseases. The current study focuses mainly on two members of the ADAMTS family, ADAMTS13 and ADAMTS7. Project 1 will investigate the allosteric regulation of ADAMTS13, an important regulator of blood clotting. I recently discovered that ADAMTS13 adopts a quiescent closed conformation and becomes allosterically activated by its substrate, Von Willebrand Factor (VWF). This proposal outlines a mechanistic approach to study how allosteric regulation is maintained within ADAMTS13 and disrupted by its interaction with VWF. Leveraging functional, kinetic, structural and bioinformatic analyses, this project sets to identify the key structural elements of allosteric regulation in ADAMTS13. The outcomes from this work will improve our understanding of inherited and acquired blood clotting disorders caused by defects in ADAMTS13. I also anticipate that our investigation of ADAMTS13 will serve as a model to study the regulation of other ADAMTS proteases. Project 2 focuses on ADAMTS7, which is known to contribute to coronary artery disease by promoting atherosclerosis and inflammation. However, the substrate targets of ADAMTS7 and its biochemical properties remain poorly understood. Previous attempts to address these research questions led to conflicting reports in the literature, likely due to the poor quality of reagents and tools available to study this novel protease. To circumvent these challenges, we will utilize novel methods in proteomics, enzymology, and protein-protein interactions to delineate biochemical properties and ideal substrates for ADAMTS7. Newly identified substrates will form the basis of novel biochemical tools to investigate ADAMTS7 activity and are necessary for future research goals to screen for small molecule inhibitors of ADAMTS7 that may have translational applications. Summarily, the study of ADAMTS13 and ADAMTS7 enzyme catalysis and regulation lay a foundation for our understanding of thrombotic and bleeding disorders and cardiovascular and inflammatory diseases, and these outcomes can be applied to study other ADAMTS proteases. More importantly, the MIRA offers the principal investigator an incredible opportunity to recruit and mentor students and other trainees from diverse backgrounds.

项目概要 该 MIRA 提案旨在揭示金属蛋白酶的调控机制和底物 ADAMTS（具有 ThromboSpondin 基序的解整合素和金属蛋白酶）家族。这些蛋白酶被认为 参与许多生物过程，并且一些成员已知对重要的人类做出贡献 疾病。目前的研究主要集中在 ADAMTS 家族的两个成员 ADAMTS13 和 ADAMTS7 上。 项目 1 将研究 ADAMTS13（凝血的重要调节因子）的变构调节。我 最近发现 ADAMTS13 采用静态闭合构象并变得变构 被其底物血管性血友病因子 (VWF) 激活。该提案概述了一种机械的研究方法 ADAMTS13 内如何维持变构调节，以及如何通过其与 VWF 的相互作用来破坏变构调节。杠杆作用 功能、动力学、结构和生物信息学分析，该项目旨在确定关键的结构元素 ADAMTS13 中的变构调节。这项工作的成果将提高我们对遗传的理解 以及由 ADAMTS13 缺陷引起的获得性凝血障碍。我还预计我们的调查 ADAMTS13 将作为研究其他 ADAMTS 蛋白酶调控的模型。项目2重点关注 ADAMTS7 已知会通过促进动脉粥样硬化和 炎。然而，ADAMTS7 的底物靶点及其生化特性仍然很差 明白了。之前解决这些研究问题的尝试导致了文献中相互矛盾的报告， 可能是由于用于研究这种新型蛋白酶的试剂和工具质量较差。为了规避这些 挑战，我们将利用蛋白质组学、酶学和蛋白质-蛋白质相互作用中的新方法来描述 ADAMTS7 的生化特性和理想底物。新确定的底物将构成 研究 ADAMTS7 活性的新型生化工具，对于未来的研究目标筛选是必要的 可能具有转化应用的 ADAMTS7 小分子抑制剂。概括地说，研究 ADAMTS13 和 ADAMTS7 酶的催化和调节为我们理解血栓形成奠定了基础 和出血性疾病以及心血管和炎症性疾病，这些结果可应用于 研究其他 ADAMTS 蛋白酶。更重要的是，MIRA 为首席研究员提供了令人难以置信的 有机会招募和指导来自不同背景的学生和其他学员。